Metabolic markers for mitochondrial function

线粒体功能的代谢标志物

• 批准号: 8697051
• 负责人: Cynthia Therese McMurray
• 金额: $ 50.87万
• 依托单位: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-20 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8697051
• 关键词: 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; 1-Methyl-4-phenylpyridinium; Abbreviations; Address; Adenosine Triphosphate; Affect; Age; Amino Acids; Animal Model; Animals; Biological Markers; Biology; Blood; Blood - brain barrier anatomy; Brain; Calcium; Cells; Chronic; Chronic Disease; Clinical Markers; Complex; Cytosol; Defect; Detection; Disease; Early Diagnosis; Environment; Environmental Exposure; Exposure to; Functional disorder; Heterogeneity; Human; Individual; Injury; Intervention; Lead; Link; Mass Spectrum Analysis; Measurable; Measurement; Measures; Metabolic; Metabolic Diseases; Metabolic Marker; Methodology; Mitochondria; Modeling; Monoamine Oxidase; Nanostructures; Noise; Oxidation-Reduction; Oxidative Phosphorylation; Pattern; Preventive Intervention; Production; Property; Propionic Acids; Resolution; Respiration; Rodent; Rotenone; Sampling; Signal Transduction; Sorting - Cell Movement; Staging; Symptoms; Synaptosomes; System; Techniques; Technology; Testing; Tissues; Toxic Environmental Substances; Toxic effect; Toxicant exposure; Toxin; Ubiquitin; Urine; Variant; base; brain cell; cell motility; cell type; dopamine transporter; functional decline; in vivo; mitochondrial dysfunction; mouse model; multicatalytic endopeptidase complex; nervous system disorder; new technology; oxidative damage; prevent; response; tool

ABSTRACT Mitochondrial (MT) dysfunction is a factor in numerous chronic diseases and the toxicity related to environmental exposures, but early deficits in MT function are difficult to detect. Current clinical markers for mitochondrial dysfunction typically detect only advanced symptoms of tissue injury and disease, yet the sensitivity to detect mild MT dysfunction and heterogeneity within tissue has hampered robust identification of meaningful biomarkers at early stages. Mitochondrial biology is variable; and chronic, low level MT dysfunction may be below the detection sensitivity of many techniques. We need new tools to enhance the mechanistic understanding of environmentally-induced mitochondrial toxicity at early stages to enable prevention and intervention. To address this problem, we have developed and applied a new technology for single cell mass spectrometry, called Nanostructure-Initiator Mass Spectrometry (NIMS). NIMS has both the single cell resolution (1-10 ¿m) and the high sensitivity (attomolar) needed to detect early biomarkers of MT dysfunction as metabolic "signatures" in individual cells. NIMS offers a number of advantages over standard mass spectrometry, including (1) ultra-high sensitivity, (2) high selectivity, and (3) single cell resolution to reduce sample complexity. We apply NIMS to identify metabolic signatures for early MT dysfunction in the brain and blood of diseased animals or animals treated with environmental toxins at "subclinical" levels. In Aim 1, we use NIMS to generate metabolic signatures for MT decline. In Aim 2, we will functionally test whether the biomarker reflects functional changes in MT or MT within the context of the cell. NIMS can be applied to any tissue and any cell type, to quantitatively sort out complex changes that occur in dynamic cellular environments, and minimizes the inherent system heterogeneity that has confounded efforts in detecting meaningful markers of MT decline.

摘要 线粒体(MT)功能障碍是许多慢性疾病的一个因素，其毒性与 环境暴露，但MT功能的早期缺陷很难检测到。目前的临床标记物 线粒体功能障碍通常只能检测到组织损伤和疾病的晚期症状，但 对检测轻度MT功能障碍和组织内异质性的敏感性阻碍了对 早期阶段有意义的生物标志物。线粒体生物学是可变的；慢性、低水平MT功能障碍 可能低于许多技术的检测灵敏度。我们需要新的工具来增强机械性 在早期阶段了解环境诱导的线粒体毒性，以便能够预防和 干预。为了解决这个问题，我们开发并应用了一种新的单细胞质量技术 光谱分析，称为纳米结构引发质谱学(NIMS)。NIMS既有单细胞 分辨率(1-10？m)和高灵敏度(大气压)需要检测MT功能障碍的早期生物标志物 作为个体细胞的新陈代谢“信号”。与标准质量相比，NIMS具有许多优势 光谱学，包括(1)超高灵敏度，(2)高选择性，(3)降低单细胞分辨率 样本复杂性。我们应用NIMS来识别大脑和大脑中早期MT功能障碍的代谢特征 患病动物的血液或用“亚临床”水平的环境毒素处理的动物的血液。在目标1中，我们使用 NIMS将为MT下降生成代谢特征。在目标2中，我们将从功能上测试生物标记物 反映MT或MT在细胞环境中的功能变化。NIMS可以应用于任何组织和 任何细胞类型，以定量地分类动态蜂窝环境中发生的复杂变化，以及 最大限度地减少固有的系统异质性，这种异质性阻碍了检测有意义的标记的努力 Mt下降。